Biochemical Investigation of Pikromycin Biosynthesis Employing Native Penta- and Hexaketide Chain Elongation Intermediates
- 19 May 2005
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 127 (23) , 8441-8452
- https://doi.org/10.1021/ja042592h
Abstract
The unique ability of the pikromycin (Pik) polyketide synthase to generate 12- and 14-membered ring macrolactones presents an opportunity to explore the fundamental processes underlying polyketide synthesis, specifically the mechanistic details of chain extension, keto group processing, acyl chain release, and macrocyclization. We have synthesized the natural pentaketide and hexaketide chain elongation intermediates as N-acetyl cysteamine (NAC) thioesters and have used them as substrates for in vitro conversions with engineered PikAIII+TE and in combination with native PikAIII (module 5) and PikAIV (module 6) multifunctional proteins. This investigation demonstrates directly the remarkable ability of these monomodules to catalyze one or two chain extension reactions, keto group processing steps, acyl-ACP release, and cyclization to generate 10-deoxymethynolide and narbonolide. The results reveal the enormous preference of Pik monomodules for their natural polyketide substrates and provide an important comparative analysis with previous studies using unnatural diketide NAC thioester substrates.Keywords
This publication has 21 references indexed in Scilit:
- Precursor-Directed Biosynthesis of Epothilone in Escherichia coliJournal of the American Chemical Society, 2004
- Clinical efficacy of ketolides in the treatment of respiratory tract infectionsJournal of Antimicrobial Chemotherapy, 2004
- Epothilone C Macrolactonization and Hydrolysis Are Catalyzed by the Isolated Thioesterase Domain of Epothilone Polyketide SynthaseJournal of the American Chemical Society, 2003
- Engineering the aveC gene to enhance the ratio of doramectin to its CHC‐B2 analogue produced in Streptomyces avermitilisBiotechnology & Bioengineering, 2003
- Molecular recognition of diketide substrates by a β-ketoacyl-acyl carrier protein synthase domain within a bimodular polyketide synthaseChemistry & Biology, 1997
- Synthesis of β-keto and α,β-unsaturated N-acetylcysteamine thioestersBioorganic & Medicinal Chemistry Letters, 1995
- Diene-dienophile hydrogen-bonding control of Diels-Alder reactions with dienes bearing a remote stereogenic center. Amplification of the receptor nature of dienes by conformational tuning leading to high diastereofacial discriminationJournal of the American Chemical Society, 1991
- Modular Organization of Genes Required for Complex Polyketide BiosynthesisScience, 1991
- Cleavage of DNA by the 1,10-phenanthroline-copper ion complex. Superoxide mediates the reaction dependent on NADH and hydrogen peroxideJournal of the American Chemical Society, 1981
- Rapid chromatographic technique for preparative separations with moderate resolutionThe Journal of Organic Chemistry, 1978